DRUG CARRIER SYSTEM:

Liposomes
Introduction:
When phospholipids are dispersed in water, they spontaneously form closed structure with
internal aqueous environment bounded by phospholipid bilayer membranes, this vesicular
system is called as liposome.
Most of theraprutic molecules are unsuccessful in the developmental process due to poor
correlation between in vitro and in vivo results. Potent drugs with a narrow therapeutic index
generally require a targeted or site specific delivery. This can be achieved by using a carrier mediated
drug delivery system which includes liposomes, microparticles, nanoparticles, etc. Liposomes are closed
concentric vesicles of phospholipids in the colloidal size range of 0.01-5.0 μm. Phospholipids are
amphiphilic molecules, containing hydrophilic and hydrophobic parts which when dispersed in water,
form into bilayered vesicles, where non polar part is shielded with polar part.
Liposomes were first reported by A. D. Bangham in early 1960s, who investigated the formation of
spherical structures when phospholipids come into contact with water.

Schematic drawing of liposomes structure and lipophilic or hydrophilic drug entrapment models.

Types of Phospholipids:

Following are its types(structures):

 Phosphatidic acid (phosphatidate) (PA)

 Phosphatidylethanolamine (cephalin) (PE)
 Phosphatidylcholine (lecithin) (PC)
 Phosphatidylserine (PS)
 Phosphoinositides: Phosphatidylinositol (PI) Phosphatidylinositol phosphate (PIP)
Phosphatidylinositol bisphosphate (PIP2)
Advantages:
 Suitable for delivery of hydrophobic,
hydrophilic and amphipatic drugs and
agents
 Use for both active and passive targeting.
 Chemically and physically well
characterized entities
 Biocompatible
 Use as carrier for suitable for controlled
release drug delivery.
 Suitable to give localized action in
particular tissues.
 Suitable to administer via various routes
 Increased efficacy and therapeutic index.
 Reduction on toxicity of the encapsulation
agent.
 Improved pharmacokinetic properties.
 Can be made into Variety of drug.
 Minimum antigenicity.
Disadvantages:
 their rapid clearance from circulation due
to uptake,
 by the reticuloendothelial system(RES),
primarily in the liver
 Leakage of encaptulation drug delivery
during storage.
 Batch to batch variation.
 Once administered, can’t removed.
 Difficult in large scale manufacture and
sterilization.
 Physical /chemical stabillity
 Very high production cost
 Possibility of dumping due to faulty
administration

Types Of Liposomes:

It is based upon:-

1. Size
2. Composition
3. Conventional liposome
4. Specialty liposome

1. SIZE:

VESICLE TYPE

Unilamellar vesicle
Small unilamellar
vesicle
Medium unilamellar
vesicle
Large unilamellar
vesicle
Giant unilamellar
vesicle
Oligomellar vesicle
Multilamellar vesicle
Multi vesicular vesicle
2. COMPOSITION:

Conventional liposome- it is prepared by normal way. Phospholipid is either neutral or negatively

charged. It is used for reticoendothelial system.

pH sensitive liposome-it is based on the passive targeting. For example; in cancer pH is dropped of that
specific area.

Cationic liposome - it is based on the active targeting by applying magnetic field.

Long circulatory liposome - it is also known as STELETH LIPOSOME. It is long circulating because it
is not detectable by opsonins, so not targeted by RES.

Immunoliposome- it is used for the immune booster. Many antibodies can be attached to it.

3. BASED UPON CONVENTIONAL LIPOSOME:

1. Synthetic identical chain phospholipid

2. Glycolipids containing phospholipids

4. BASED UPON SPECIALTY LIPOSOME:

1. Bipolar fatty acid

2. Antibody detected liposome
3. Lipoprotein coated liposome
4. Carbohydrate coated liposome
5. Multiple encapsulated liposome

Methods of preparation:

1. Active loading If drug is added after the formation of liposome it is called active loading.
2. Passive loading: During the formation of liposome, if the drug is added this is called passive
loading.

Basic concept/ fundamental steps:

1. Drying step:

Phospholipids solubilize in organic solvent. Organic solvent evaporates and drying occurs. Organic
solvent added so that a thin layer can be formed and for the uniform mixing of cholesterol, drug and
phospholipid.

2. Hydration:
This mixture is added slowly into water or the water is added into it slowly. Phospholipid orient
themselves and convert into liposomes.

3. Purification, analysis and sizing:

1. MECHANICAL DISPERSION METHOD by passive loading:
Hydration of a Thin Lipid Film: Bangham Method.
A mixture of phospholipid and cholesterol were dispersed in organic solvent. Then, the organic solvent
was removed by means of evaporation (using a Rotary Evaporator at reduced pressure). Finally, the dry
lipidic film deposited on the flask wall was hydrated by adding an aqueous buffer solution under agitation
at temperature above the lipid transition temperature.
Reverse-Phase Evaporation (REV) Technique.
A lipidic film is prepared by evaporating organic solvent under reduced pressure. The system is purged
with nitrogen and the lipids are re-dissolved in a second organic phase which is usually constituted by
diethyl ether and/or isopropyl ether. The organic solvent is subsequently removed and the system is
maintained under continuous nitrogen.
Ultrasonic Method
Ultrasonication of an aqueous dispersion of phospholipids is done by two types of sonicators i.e. either
probe Sonicators(used for the small volume which requires high energy )or bath sonicators(employed for
the large).
2. SOLVENT (Ether or Ethanol) DISPERSION METHOD
The solvent injection methods involve the dissolution of the lipid into an organic phase (ethanol or ether),
followed by the injection of the lipid solution into aqueous media, forming liposomes.
In ethanol injection method, the lipid is injected rapidly through a fine needle into an excess of saline or
other aqueous medium. In ether injection method the lipid is injected very slowly through a fine needle
into an excess of saline or other aqueous medium.
3. DETERGENT REMOVAL METHID
The detergents at their critical micelles concentrations have been used to solubilize lipids. As the
detergent is removed the micelles become progressively richer in phospholipid and finally combine to
form LUVs. The detergents can be removed by dialysis. The advantages of detergent dialysis method are
excellent reproducibility and production of liposome populations which are homogenous in size. The
main drawback of the method is the retention of traces of detergent(s) within the liposomes.
Proliposomes
In proliposomes, lipid and drug are coated onto a soluble carrier to form free-flowing granular material
which on hydration forms an isotonic liposomal suspension. The proliposome approach may provide an
opportunity for cost-effective large scale manufacturing of liposomes containing particularly lipophilic
drugs.
Limitations of Liposomes
Liposomes have shown a great potential as carrier based drug delivery system. However, some of the
limitations associated with the development of liposomes are product stability, low drug entrapment, short
circulation half-life of liposomes, vesicle size and high production costs etc. The stability of the
liposomes is majorly governed by the final composition of the liposomal formulation. Physical instability
occurs predominantly due to oxidation of unsaturated acyl chains or hydrolysis of ester bonds, drug
leaking and aggregation of vesicles resulting in bigger liposomes which are rapidly cleared by
Reticuloendothelial system (RES).
Characterization Of Liposomes:

After preparation and before use, the liposome must be characterized. Evaluation could be classified into
three broad categories which are physical(size, morphology, lamellarity and shape) , chemical (potency
and purity)and biological methods.
Vesicle Size
The average size and size distribution of liposomes are important parameters especially when the
liposomes are intended for therapeutic use by inhalation or parenteral route. Various methods for
determining size and size distribution(microscopy techniques, size-exclusion chromatography etc) are
available.
Encapsulation Efficiency: Or encapsulation capacity, define as the total amount of encapsulant found in
the liposome solution versus the total initial input of encapsulant solution. The liposome preparations are
a mixture of encapsulated and un-encapsulated drug fractions. The first step for the determination of the
encapsulation efficiency is the separation between the encapsulated drug (within the carrier) and the free
drug.Several separation techniques available mini-column centrifugation, dialysis membrane,
ultracentrifugation technique etc.

Zeta Potential The zeta potential of a particle is the overall charge that a particle acquires in a particular
medium. It is a physical property which is exhibited by any particle in suspension. Measurements of zeta
potential are commonly used to predict the stability of colloidal systems. Suspension with zeta potentials
> +30 mV or < −30 mV are normally considered stable.

Therapeutic Applications:
1. Formulation aid
Hydrophobic drugs such as cyclosporin and paclitaxel are usually formulated in surfact ants and organic
co-solvents for systemic administration in humans. These solubilizers may cause toxicity at the doses
needed to deliver the drug. In contrast, liposomes are made up of lipids which are relatively non-toxic,
non-immunogenic, biocompatible and biodegradable molecules, and can encapsulate a broad range of
water-insoluble (lipophilic) drugs.
2. Intracellular drug delivery
Drugs with intracellular targets/receptors are required to cross the plasma membrane for pharmacological
activity. Liposomal delivery of drug can be very effective because liposomes have structural similarity
with biological membrane and are predominantly taken up by the cells. For example, the potency of
PALA (N-phosphonacetyl-L-aspartate) encapsulated liposome’s was up to 500-fold greater against
human ovarian tumor cell lines than that of free PALA
3. Gene therapy
A number of systemic diseases are caused by lack of enzymes/factors which are due to missing or
defective genes. Cationic liposomes systems can be used for the delivery of such missing/defected gene
(plasmid). They are usually composed of a cationic lipid derivative and a neutral phospholipid. The
negatively charged genetic material (e.g., plasmid) is not encapsulated in liposomes but complexed with
cationic lipids by electrostatic interactions. These complexes are thought to enter the cell by fusion with
the plasma or endosome membrane.
4. Site-avoidance delivery
Drugs used in the treatment of diseases like cancer usually have a narrow therapeutic index (TI) and can
be highly toxic to normal tissues. Liposomes are taken up poorly by tissues such as heart, kidney, and GI
tract, which are major sites for toxic side-effects of a variety of anti neoplastic drugs. Thus, liposome
formulation may improve the TI by altering the bio distribution of drug away from drug sensitive normal
tissues.
5. Site-specific targeting
Site-specific delivery, the concept involves the delivery of a larger fraction of drug to the target site and
therefore, reducing exposure to normal tissues. Liposomes have been employed for accomplishing both
passive and active targeting of drugs.
Passive targeting
This approach for liposome drug delivery exploits the natural tendency of certain cells such as Kupffer
cells in the liver, and circulating macrophages of RES to phagocytose foreign microparticulates such as
liposomes.
Active targeting
Active targeting of liposome encapsulated drugs may be accomplished by coupling specific antibodies to
vesicles (immune liposomes). Long circulating immunoliposomes (hydrophilic polymer- coated vesicles
bound to antibodies and <0.15 micron in size) can now be designed which may be able to recognize and
bind with greater specificity to target cells following systemic administration.
6. Intra peritoneaI administration
Direct administration of antineoplastic agents into the intraperitoneal (i.p.) cavity is therapeutically
advantageous for cancers but it is unsuccessful for free drugs because of relatively fast clearance of the
drugs from the i.p. cavity resulting in lowered concentrations at the site of action. However, the clearance
of liposomes from the peritoneal cavity is significantly slower than that of free drug and therefore, higher
drug concentrations can be achieved for extended periods of time with the use of liposome formulations.
7. Immunological adjuvant in vaccines
Liposomes can encapsulate antigens in their aqueous space or incorporate in the bilayer depending on the
lipophilicity of the antigen. Liposome also have been used as nontoxic adjuvants with bacterial, viral,
protozoan, tumor and other antigens
References:

V. Ravichandiran, K. Masilamani, B. Senthilnathan Pelagia Research Library Der Pharmacia Sinica,

2011, 2 (1): 19-30
Abdus Samad, Y. Sultana and M. Aqil, Liposomal Drug Delivery Systems: An Update Review

R. BANERJEE, Cardiovascular Research Institute, 3333 California Street, Suite 150, University of
California, San Francisco, CA 94118-1245 Liposomes: Applications in Medicine

Vijaykumar Nekkant, Sandeep Kalepu, Pharmaceutical Nanotechnology, 2015, 3, 35-55 35 Recent

Advances in Liposomal Drug Delivery: A Review